PHOSPHATIDYL-L-SERINE AND PHOSPHATIDIC ACID COMPOSITION AS ANTI-DEPRESSANT, STRESS SUPPRESSOR AND MOOD IMPROVER

COMPOSITION ACIDE PHOSPHATIDIQUE ET DE PHOSPHATYIDYL-L-SERINE COMME ANTIDEPRESSEUR, REDUCTEUR DE STRESS ET PSYCHOTROPE

English Abstract

An anti-depressant, mental & emotional stress suppressor and mood improver having a prominent action for decreasing blood cortisol level and serotonin reuptake and has an effect of alleviating symptoms associated with depression and mental & emotional stress of a subject administered with the improver. The improver contains as the effective ingredient a combination of phosphatidyl-L-serine and phosphatidic acid, or the salts thereof, comprising at least 200 (w/w) phosphatidyl-L-serine and typically within the range of about 20%-40% of phosphatidyl-L-serine, out of the total phospholipid content of the composition and at least 3% (w/w) of phosphatidic acid, preferably above about 10% and typically within the range of about 20%-40% of phosphatidic acid, out of the total phospholipid content of the composition. The phosphatidyl-L-serine and phosphatidic acid has a structural fatty acid chain derived from at least one raw material lecithin selected from the group consisting of soy bean lecithin, rapeseed lecithin or egg yolk lecithin. Using the raw material lecithin as the substrate, phosphatidyl-L-serine and phosphatidic acid can be produced by enzymatic conversion utilizing phospholipase-D.

French Abstract

Un agent antidépresseur, suppresseur de stress émotionnel et mental, et améliorateur de l'humeur, dont l'action prédominante est d'abaisser le niveau de cortisol sanguin et la réabsorption de la sérotonine, et qui soulage les symptômes associés avec la dépression et le stress mental et émotionnel chez un sujet ayant reçu cet agent d'amélioration. L'agent d'amélioration contient comme ingrédient actif un mélange de phosphatidyl-L-sérine et d'acide phosphatidique, ou de leurs sels, comprenant au moins 20 % (en poids) de phosphatidyl-L-sérine et typiquement, environ entre 20 %-40 % de phosphatidyl-L-sérine, sur la concentration totale en phospholipides du mélange et au moins 3 % (en poids) d'acide phosphatidique, de préférence plus de 10 % environ, et typiquement entre environ 20 %-40 % d'acide phosphatidique, sur la concentration totale en phospholipides du mélange. La phosphatidyl-L-sérine et l'acide phosphatidique présentent une chaîne d'acide gras structurale dérivée d'au moins une lécithine en tant que matière première provenant du groupe comprenant la lécithine de soja, la lécithine de colza ou la lécithine de jaune d'oeuf. En utilisant cette matière première de lécithine comme substrat, la phosphatidyl-L-sérine et l'acide phosphatidique peuvent être produits par conversion enzymatique utilisant une phospholipase D.

Claims

CLAIMS:
1. An anti-depressant, mental and emotional stress
suppressor, and mood improver composition, comprising at
least 20% (w/w) phosphatidyl-L-serine and at least 3% (w/w)
phosphatidic acid or salts thereof, as the effective
ingredient, out of the total phospholipid content of the
composition, wherein the phosphatidyl-L-serine and
phosphatidic acid have a structural fatty acid chain derived
from at least one raw material lecithin selected from the
group consisting of soy bean lecithin, rapeseed lecithin and
egg yolk lecithin, and which is produced by reaction with
phospholipase-D.
2. The composition of claim 1, comprising from about
20% to 40% (w/w) phosphatidyl-L-serine or a salt thereof.
3. The composition of claim 1 or 2, comprising above
about l0% (w/w) phosphatidic acid or a salt thereof.
4. The composition of claim 3, comprising about 20%
to about 40% (w/w) phosphatidic acid or a salt thereof.
5. An antidepressant, mental and emotional stress
suppressor, and mood improver composition, comprising
phosphatidyl-L-serine and phosphatidic acid or salts
thereof, as the effective ingredient, wherein the
phosphatidyl-L-serine and the phosphatidic acid have a
structural fatty acid chain derived from at least one raw
material lecithin selected from the group consisting of soy
bean lecithin, rapeseed lecithin and egg yolk lecithin, and
which is produced by enzymatic reaction with phospholipase-
D, and wherein the structural fatty acid chain is a
hydrogenated saturated fatty acid chain.
6

6. The composition of any one of claims 1 to 5,
further comprising a pharmaceutically acceptable or food
excipient.
7. Use of a composition of any one of claims 1 to 6,
for improving symptoms of depression, mental and emotional
stress, and mood in a subject in need thereof.
8. Use of a composition of any one of claims 1 to 6,
for improving symptoms of depression, mental and emotional
stress, and mood in a subject in need thereof by reducing
the blood cortisol level and serotonin reuptake in the brain
of the subject.
9. Use of a composition of any one of claims 1 to 6,
for preparing a medicament for improving symptoms of
depression, mental and emotional stress, and mood in a
subject in need thereof.
10. Use of a composition of any one of claims 1 to 6,
for preparing a medicament for improving symptoms of
depression, mental and emotional stress, and mood in a
subject in need thereof by reducing the blood cortisol level
and serotonin reuptake in the brain of the subject.
11. A commercial package comprising a composition of
any one of claims 1 to 6, and associated therewith
instructions for the use thereof in improving symptoms of
depression, mental and emotional stress, and mood in a
subject in need thereof.
12. A commercial package comprising a composition of
any one of claims 1 to 6, and associated therewith
instructions for the use thereof in improving symptoms of
7

depression, mental and emotional stress, and mood in a
subject in need thereof by reducing the blood cortisol level
and serotonin reuptake in the brain of the subject.
8

Description

CA 02352528 2005-03-02
28900-53
DESCRIPTION
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a composition having an effect of alleviating
symptoms
associated with depression and mental and emotional stress.
2. Description of the Related Art
A. Zanotti et al. report that the oral administration of phosphatidylserine
extracted from
bovine brain to aged rats with memory deficits for 12 weeks improved the
performance of
the aged rats (A. Zanotti et al., Psychopharmacology Berl., Vol. 99, P. 316,
1989).
Monteleoni et al., (Eur. J. Clin. Pharmacology, 385-388, 1992) investigated
the chronic administration of
brain cortex phosphatidylserine on the neuroendocrine responses to physical
stress. The study showed that
oral administration of phosphatidylserine at 800 mg per day for 10 days prior
to exercise, reduced the ACTH
and cortisol responses to physical exercise. A 400 mg per day dose was shown
to produce no effect on the
cortisol response.
Monteleoni et al: further reported (Neuroendocrinology, 52, 243-248, 1990) on
the influence of brain cortex
phosphatidylserine on the neuroendocrine and neurovegetative responses to
physical stress. In a double blind
study, every participant received intravenously brain cortex
phosphatidylserine or a placebo before starting a
physical exercise. Blood samples were collected before and after the exercise
for plasma ACTH, cortisol and
growth hormone readings. It outcome showed that in the placebo group the
physical stress induced an
increase in ACTH, cortisol and growth hormone while the phosphatidylserine
group showed a reduction in
production of ACTH and cortisol.
In a series of patents (PCT No. PCT/IL97100147 Sec. 371 Date Feb. 24, 1999
Sec. 102(e)
Date Feb. 24, 1999 PCT Filed May 6, 1997 PCT Pub. No. W097/41874 PCT
Pub. Date Nov. 13, 1997, Priority Number(s): IL19960118180 19960508;
W01997IL00147 19970506) phosphatidic acid has been shown to alleviate
withdrawal
symptoms associated with addiction (cigarettes, alcohol, narcotics).
SUMMARY OF THE INVENTION
The present invention provides an improver having an effect of alleviating
symptoms
associated with depression and mental and emotional stress.
According to the research work of the present inventors, it is confirmed that
a complex of
phosphatidyl-L-serine and phosphatidic acid produced by an enzymatic
conversion utilizing
phospholipase-D of at least one raw material lecithin selected from the group
consisting of
soybean lecithin, rapeseed lecithin, and egg yolk lecithin, has in mental and
emotional
1

CA 02352528 2005-03-02
28900-53
stress and depression a prominent effects of decreasing
cortisol blood level and serotonin uptake to normal level.
In one aspect, the invention provides an anti-depressant,
mental and emotional stress suppressor, and mood improver
composition, comprising at least 200 (w/w) phosphatidyl-L-
serine and at least 30 (w/w) phosphatidic acid or salts
thereof, as the effective ingredient, out of the total
phospholipid content of the composition, wherein the
phosphatidyl-L-serine and phosphatidic acid have a
structural fatty acid chain derived from at least one raw
material lecithin selected from the group consisting of soy
bean lecithin, rapeseed lecithin and egg yolk lecithin, and
which is produced by reaction with phospholipase-D.
Typically, the composition comprises about 20o to 400 (w/w)
phosphatidyl-L-serine or a salt thereof and above 100,
preferably about 20% to 40% (w/w), phosphatidic acid or a
salt thereof.
In a further aspect, the invention provides an
antidepressant, mental and emotional stress suppressor, and
mood improver composition, comprising phosphatidyl-L-serine
and phosphatidic acid or salts thereof, as the effective
ingredient, wherein the phosphatidyl-L-serine and the
phosphatidic acid have a structural fatty acid chain derived
from at least one raw material lecithin selected from the
group consisting of soy bean lecithin, rapeseed lecithin and
egg yolk lecithin, and which is produced by enzymatic
reaction with phospholipase-D, and wherein the structural
fatty acid chain is a hydrogenated saturated fatty acid
chain.
The compositions of the invention may further comprise a
pharmaceutically acceptable or food excipient, e.g., as
2

CA 02352528 2005-03-02
28900-53
detailed in "Remington's Pharmaceutical Sciences", Mack
Publishing Co., Easton, PA, USA, 19th Edition, 1995.
The invention also provides uses of the compositions of the
invention for improving symptoms of depression, mental and
emotional stress, and mood in a subject in need thereof and
for preparing a medicament for such use.
The invention also provides a commercial package comprising
a composition of the invention and associated therewith
instructions for the use thereof in improving symptoms of
depression, mental and emotional stress, and mood in a
subject in need thereof.
The compositions of the invention reduce blood cortisol
level and serotonin reuptake in the brain.
An improver of the present invention contains phosphatidyl-
L-serine and phosphatidic acid or the salt thereof as the
effective ingredient, wherein the phosphatidyl-L-serine and
phosphatidic acid have a structural fatty acid chain derived
from at least one raw material lecithin selected from the
group consisting of soy bean lecithin, rapeseed lecithin,
and egg yolk lecithin.
The improver of the present invention may be administered
effectively via intravenous administration and oral
administration. The improver may be mixed with other
excipients such as additional phospholipids and lyso-
phospholipids, sugar and protein to prepare capsules and
granules with improved handling and shelf life. Because of
the absence of any safety problem, the improver may be
blended into daily foods and beverages, either in powder or
liquid form or as hydrogenated substance for use in
2a

CA 02352528 2005-03-02
28900-53
improving and preventing mental and emotional stress and
depression symptoms.
The aforementioned phosphatidyl-L-serine and phosphatidic
acid as the effective ingredients in accordance with the
present invention are both produced by the enzymatic
reaction with phospholipase-D using as the substrate soy
bean lecithin, rapeseed lecithin or egg yolk lecithin.
The process will now be illustrated. A raw material
lecithin (namely, phosphatidylcholine) selected from soy
bean lecithin, rapeseed lecithin and egg yolk lecithin is
subjected to the process of transphosphatidylation and
hydrolysis with phospholipase-D in the presence of L-serine
and water, thereby substituting the choline group with the
serine group or the hydroxyl group, to produce the
rearranged phosphatidyl-L-serine and phosphatidic acid.
Any commercially available soy bean lecithin, rapeseed
lecithin or egg yolk lecithin may be used, with no
limitation, as the raw material. As phospholipase-D for use
in the process of enzymatic conversion, use may be made of
for example those from cabbage and actinomyces, if they have
an activity on lecithin or hydrogenated lecithin or
lysolecithin in the presence of L-serine and water to
produce phosphatidyl-L-serine and phosphatidic acid.
A specific process of enzymatic conversion is known and
described in for example the article by Eibl A. and
Kovatchev S. "Preparation of phospholipids analogs by
phospholipase-D". ("Methods in Enzymology" Vol. 72, pages:
632-639, 1981), so no detailed explanation is described
herein.
2b

CA 02352528 2001-07-06
DESCRIPTION OF THE PREFERRED EMBODIMENTS
EXAMPLE 1-1
Using soy bean lecithin as the raw material, phosphatidyl-L-serine and
phosphatidic acid
were produced by the following process.
Soy bean lecithin (50 g; Epikuron 135 as the product name; Lucas Meyer GmbH,
Germany) and soybean oil ( 10 g) were placed in a 300-ml vial, followed by
addition of
ethyl acetate (50 ml) for solubilization. Adding a solution (20 ml) of 0.30
g/ml L-serine
dissolved in 0.1 M sodium phosphate buffer, pH 7.0 to the resulting solution
for thorough
blending, a solution of 500 U/ml phospholipase-D from cabbage was added to the
mixture
solution for reaction at 25. degrees C. for 5 hours under stirring with a
stirrer.
So as to inactive the enzyme in the reaction solution, the vial containing the
reaction
solution was immersed in hot water. Subsequently, the reaction solution was
cooled in ice
to separate the solution into two layers, which were then left to stand for 30
minutes.
Subsequently, the upper layer was discarded. The remaining lower layer was
extracted in
chloroform, which was then dried under reduced pressure.
EXAMPLE 1-2
Using egg yolk lecithin (DS-PL95E as the product name; manufactured by Doosan
Corp.
Venture BG Biotech BU. Korea) as the substrate, rearranged phosphatidyl-L-
serine and
phosphatidic acid were produced by the same method as in Example 1-1.
EXAMPLE 2-1
Soy bean lecithin (Epikuron 135 as the product name; Lucas Meyer GmbH,
Germany) was
processed for hydrogenation. Using the hydrogenated soy bean lecithin as the
substrate,
phosphatidyl-L-serine and phosphatidic acid were produced by the same method
as in
Example 1-1.
EXAMPLE 2-2
The soy bean lecithin-derived phosphatidyl-L-serine and phosphatidic acid were
(1 g)
produced in Example 1-l, was solubilized in a mixture solution of n-hexane (IS
g) and
ethanol (3 g). Adding 10% palladium carbon (0.1 S g) to the solution, the
resulting solution
was processed for hydrogenation for about 5 hours under stirring under the
conditions of
room temperature and ambient pressure.
EXAMPLE 3
As will be described below, the effect of improving mood and depression level
via oral
administration was confirmed at a test. phosphatidyl-L-serine and phosphatidic
acid
complex at a ratio of 1:1 (M/M) was prepared by Lipogen Products (9000) Ltd.
via the
process of enzymatic reaction from a substrate soy bean lecithin according to
Example 1-1.
3

CA 02352528 2001-07-06
Five volunteers with depression symptoms received 200 mg. three times per day
for a
period of four weeks. The results are presented in the following table 1.
Table I
Subject Age Depression Depression
symptoms before*symptoms after*
Male 50 ++ +
Male 61 ++ 0
Female 53 + 0
Female 35 ++ +
Female 42 ++ +
* 0 - Normal,
+ Slightly depressed.
++ Highly depressed.
The depression scale was assessed by the subject.
As indicated in Table l, a significant improvement was observed in all five
participants
irrespective of age or gender.
EXAMPLE 4
As will be described below, the effect of reduction of blood cortisol level
under mental and
emotional stress via oral administration was confirmed at a test. phosphatidyl-
L-serine and
phosphatidic acid complex at a ratio of I:l (M/M) was prepared by Lipogen
Products
(9000) Ltd. via the process of enzymatic reaction from a substrate soy bean
lecithin
according to Example 1-1. Four student volunteers in a period of critical
examination with
stress symptoms received 200 mg. three times per day for a period of one week.
The results
are presented in the following table 2.
Table 2
Subject Age Blood cortisol Blood cortisol
level before level
(nM). after (nM).
Male 24 10 2
Male 26 22 8
Female 23 18 4
Female 35 12 8
Remark: The cortisol level was measured by convent~ona~ enzyme ~mmuno assay.
As indicated in Table 2, the cortisol level was significant reduced and
approached normal
level. improvement was observed in all four participants irrespective of age
or gender.
EXAMPLE 5
4

CA 02352528 2001-07-06
As will be described below, the effect of phosphatidyl-L-serine and
phosphatidic acid
complex at a ratio of l:l (M/M) on serotonin uptake after oral administration
was
contirmed at a test. phosphatidyl-L-serine and phosphatidic acid complex at a
ratio of I:1
(M/M) was prepared by Lipogen Products (9000) Ltd. via the process of
enzymatic reaction
from a substrate soy bean lecithin according to Example 1-1. Five volunteers
with
depression symptoms received 200 mg. three times per day for a period of four
weeks.
Peripheral blood samples obtained from the subjects presented in Example 3,
were
separated to leucocytes, and serotonin uptake was measured with tritiated
serotonin as
described by Faraj et al. (Int. J. Immunopharm. 16, 561-567, 1994). The
results are
presented in the following table 3.
Table 3
Subject Age Change in
sertonin uptake
level
Male 50 -40%
Male 61 -20%
Female 53 -15%
Female 35 -55%
Female 42 -58%
As indicated in Table 3, the sertonin uptake level was significant reduced due
to
the treatment with the improver in all five participants irrespective of age
or gender.
The results with the leucocytes represents the expected changes in the brain
(Faraj et al.
Pharmacology. 42, 135-141, 1991). Such a reduction in serotonin uptake can be
implicated
in various positive aspects of mood improvement (Owens & Nemeroff, Clin. Chem.
40,
288-295, 1994).
As has been described above, the anti-depressant, mental & emotional stress
suppressor and
mood improver containing phosphatidyl-L-serine and phosphatidic acid from soy
bean,
rapeseed or egg yolk as the effective ingredient in accordance with the
present invention
can be continuously administered readily with no pain because phosphatidyl-L-
serine and
phosphatidic acid effective for improving and alleviating symptoms associated
with
depression and mental & emotional stress can be orally ingested from the
improver.
Furthermore, the phosphatidyl-L-serine and phosphatidic acid effective for
improving and
alleviating symptoms associated with depression and mental & emotional stress
can be
produced at less cost and additionally at a large scale, by utilizing
enzymatic conversion via
a phospholipid degradation enzyme (phospholipase-D).
*****